Protective enzymes

ABSTRACT

The present invention relates to methods and compositions that include enzymes and/or binding polypeptides useful for protecting polymers from damage caused by fatty acids from secreted biological fluids such as sebum or sweat.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application claims priority to U.S. Provisional ApplicationNo. 62/557,544, filed Sep. 12, 2017, the content of which is hereinincorporated by reference in its entirety.

SEQUENCE LISTING

The present specification makes reference to a Sequence Listing(submitted electronically as a .txt file named “Sequence Listing 0174”.The .txt file was generated on Sep. 12, 2018, and is 73,728 bytes insize. The entire content of the Sequence Listing is herein incorporatedby reference.

BACKGROUND

Polymeric materials may be vulnerable to degradation from contaminants.

SUMMARY

The present disclosure encompasses the recognition that certain enzymesand polypeptides may be useful to neutralize (e.g., prevent or mitigate)damage to polymeric compositions caused by lipids. Exposure to certainsecreted biological fluids containing lipids (e.g., human sweat and skinoils) can cause some polymers to swell and degrade. In some embodiments,an enzyme of the present disclosure facilitates degradation of a lipid,and thereby prevents damage to a polymeric structure. In someembodiments, an enzyme useful in the context of the present disclosurefacilitates degradation of harmful lipids such as fatty acids.

The present disclosure provides methods and compositions that protectcomponents of a composition (e.g., polymers) from damage by lipids(e.g., fatty acids) and/or other compounds that may cause damage. Insome embodiments, a lipid and/or other compound that may cause damage ispresent in a biological fluid. In some embodiments methods andcompositions are provided that include protective enzymes useful forbreakdown of components in biological fluids that may cause damage to apolymer. In some embodiments, methods and compositions are providedwhere one or more protective enzymes are embedded within a composition.In some embodiments, methods and compositions are provided where one ormore protective enzyme are applied to the surface of a composition(e.g., as a coating).

In some embodiments, methods are provided for promoting breakdown of oneor more lipids (e.g., fatty acids) in a biological fluid, the methodcomprising contacting the biological fluid with an enzyme. In someembodiments, a method for promoting breakdown of one or more lipids in abiological fluid comprises contacting the biological fluid with aplurality of enzymes.

In some embodiments, methods are provided for inhibiting swelling of apolymer caused by exposure to a fatty acid, the methods comprisingcontacting the polymer with an enzyme. In some embodiments, methodscomprise contacting the polymer with a plurality of enzymes. In someembodiments, methods comprise embedding a plurality of enzymes within apolymer. In some embodiments, methods comprise coating a polymericcomposition with a plurality of enzymes.

In some embodiments, methods are provided for inhibiting swelling of apolymer caused by exposure to a fatty acid. The methods comprisecontacting the polymer with a polypeptide that binds the fatty acid. Insome embodiments, a polypeptide forms a complex with the fatty acid. Insome embodiments, a polypeptide binding to a fatty acid inhibitsreactivity of the fatty acid. In some embodiments, a polypeptide bindsto a fatty acid, and thereby prevents it from damaging a polymer. Insome embodiments, a polypeptide inhibits diffusion of the fatty acid.

In some embodiments, compositions are provided comprising an enzymeembedded in a polymer. In some embodiments, compositions comprise two ormore enzymes embedded in a polymer. In some embodiments, compositionscomprise a polymer and two or more enzymes that promote breakdown of afatty acid.

In some embodiments, compositions are provided comprising a polymer andone or more enzymes that promote breakdown of a fatty acid and one ormore polypeptides that form a complex with a fatty acid. In someembodiments, a polypeptide binding to a fatty acid inhibits thereactivity of the fatty acid. In some embodiments, a polypeptide bindsto a fatty acid, and thereby prevents it from damaging a polymer. Insome embodiments, a polypeptide inhibits diffusion of the fatty acid.

In some embodiments, topical formulations are provided comprising one ormore enzymes that promote breakdown of a fatty acid. In someembodiments, topical formulations are provided comprising one or morepolypeptides that form a complex with a fatty acid.

In some embodiments, compositions are provided comprising one or moreenzymes that promote breakdown of a fatty acid and one or morepolypeptides that form a complex with a fatty acid. In some embodiments,polypeptide binding to a fatty acid inhibits the reactivity of the fattyacid. In some embodiments, a polypeptide binds to a fatty acid, therebyprevent it from damaging a polymer. In some embodiments, a polypeptideinhibits diffusion of the fatty acid.

In some embodiments, a biological fluid is secreted by a human. In someembodiments, a biological fluid is or comprises human sweat. In someembodiments, a biological fluid is or comprises sebum. In someembodiments, a lipid in a biologic fluid is an unsaturated fatty acid.In some embodiments, unsaturated fatty acids include palmitoleic acid,oleic acid, myristoleic acid, linoleic acid, and/or arachidonic acid. Insome embodiments, a fatty acid is an oleic acid. In some embodiments, afatty acid is a linoleic acid. In some embodiments, a biological fluidcomprises a plurality of fatty acids to be neutralized.

In some embodiments, enzymes for use in a method or composition of thepresent disclosure include dioxygenases, monooxygenases, hemeperoxidases, P450s, and combinations and/or variants thereof. In someembodiments, an enzyme is a dioxygenase. In some embodiments, an enzymeis a monooxygenase. In some embodiments, methods comprise contacting thebodily fluid with a plurality of enzymes.

In some embodiments, an enzyme is of animal origin. In some embodiments,an enzyme is of plant origin. In some embodiments, an enzyme is offungal origin. In some embodiments, an enzyme is of bacterial origin. Insome embodiments, an enzyme is a cyanobacterial enzyme. In someembodiments, an enzyme is from archaea.

In some embodiments, an enzyme catalyzes beta or omega oxidation. Insome embodiments, an enzyme catalyzes hydroperoxidation at the 10Sand/or 12S-carbon of a fatty acid (e.g., an oleic acid). In someembodiments, an enzyme for use in the context of the present disclosuredoes not require adenosine triphosphate (ATP) for catalytic activity.

In some embodiments, an enzyme is a lipoxygenase and/or has lipoxygenaseactivity (e.g., 10S-LOX activity). In some embodiments, an enzyme foruse in the context of the present disclosure is a cyanobacterial enzymewith LOX activity (e.g., 10S-LOX activity) or variant thereof. In someembodiments, a cyanobacterial prostaglandin-endoperoxide synthase enzymeor variant thereof has LOX activity (e.g., 10S-LOX activity). In someembodiments, a cyanobacterial heme peroxide synthase enzyme or variantthereof has LOX activity (e.g., 105-LOX activity). In some embodiments,an enzyme with LOX (e.g., 10S-LOX activity) catalytic activity comprisesan amino acid sequence of any one of SEQ. ID NOs: 13-15. In someembodiments, an enzyme with LOX (e.g., 10S-LOX activity) catalyticactivity comprises an amino acid sequence that is at least about 50%(e.g., at least about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%,97%, 98%, or 99%) identical to any one of SEQ ID NOs: 13-15.

In some embodiments, an enzyme is a lipoxygenase. In some embodiments, alipoxygenase has 10S-LOX activity. In some embodiments, a 10S-LOXcomprises an amino acid sequence of any one of SEQ ID NOs: 1-12. In someembodiments, a 10S-LOX enzyme has an amino acid sequence that is atleast about 50% (e.g., at least about 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95%, 96%, 97%, 98%, or 99%) identical to any one of SEQ ID NOs:1-12. In some embodiments, a 10S-LOX comprises an amino acid sequence ofSEQ ID NO: 13. In some embodiments, a 10S-LOX comprises an amino acidsequence of SEQ ID NO: 14. In some embodiments, a 10S-LOX comprises anamino acid sequence of SEQ ID NO: 15. In some embodiments, a 10S-LOXenzyme comprises an amino acid sequence that is at least about 50%(e.g., at least about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%,97%, 98%, or 99%) identical to any one of SEQ ID NOs: 13-15. In someembodiments, an enzyme is a lipoxygenase that catalyzeshydroperoxidation at positions 9 and/or 13 of a fatty acid. In someembodiments, an enzyme is a lipoxygenase that catalyzeshydroperoxidation at positions 9S and/or 13S of a fatty acid. In someembodiments, an enzyme is a lipoxygenase that catalyzeshydroperoxidation at positions 9R and/or 13R of a fatty acid.

In some embodiments, an enzyme is embedded in a polymer. In someembodiments, an enzyme is applied to the surface of a polymer. In someembodiments, a polymer is a component of a device. In some embodiments,a device is an electronic device.

BRIEF DESCRIPTION OF THE DRAWINGS

The Drawings included herein, composed of the following Figures, are forillustration purposes only and not for limitation.

FIG. 1 depicts the chemical reaction catalyzed by LOX enzymes andreduction reaction used to generate a product detected during the enzymeassay.

FIG. 2 depicts enzyme activity data generated by assaying various LOXenzymes for activity on oleic acid and detecting the 10-HOME productshown in FIG. 1.

FIG. 3 depicts swelling of a representative adhesive upon exposure to anexemplary fatty acid, oleic acid. X-axis indicates the number of hourspost-exposure; Y-axis indicates the relative area of an adhesive disk.

FIG. 4 depicts swelling of a representative adhesive upon exposure to anexemplary fatty acid, oleic acid, using a second assay. X-axis indicatesthe number of hours post-exposure; Y-axis indicates the relative area ofan adhesive disk.

FIG. 5 is a schematic illustrating relatedness of certain enzymes for10S-LOX activity.

CERTAIN DEFINITIONS

In order that the present invention may be more readily understood,certain terms are first defined below. Additional definitions for thefollowing terms and other terms are set forth throughout thespecification.

About: The term “about”, when used herein in reference to a value,refers to a value that is similar, in context to the referenced value.In general, those skilled in the art, familiar with the context, willappreciate the relevant degree of variance encompassed by “about” inthat context. For example, in some embodiments, the term “about” mayencompass a range of values that within 25%, 20%, 19%, 18%, 17%, 16%,15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, orless of the referred value.

Comparable: As used herein, the term “comparable” refers to two or moreagents, entities, situations, sets of conditions, etc., that may not beidentical to one another but that are sufficiently similar to permitcomparison there between so that one skilled in the art will appreciatethat conclusions may reasonably be drawn based on differences orsimilarities observed. In some embodiments, comparable sets ofconditions, circumstances, individuals, or populations are characterizedby a plurality of substantially identical features and one or a smallnumber of varied features. Those of ordinary skill in the art willunderstand, in context, what degree of identity is required in any givencircumstance for two or more such agents, entities, situations, sets ofconditions, etc. to be considered comparable. For example, those ofordinary skill in the art will appreciate that sets of circumstances,individuals, or populations are comparable to one another whencharacterized by a sufficient number and type of substantially identicalfeatures to warrant a reasonable conclusion that differences in resultsobtained or phenomena observed under or with different sets ofcircumstances, individuals, or populations are caused by or indicativeof the variation in those features that are varied.

Designed: As used herein, the term “designed” refers to an agent (i)whose structure is or was selected by the hand of man; (ii) that isproduced by a process requiring the hand of man; and/or (iii) that isdistinct from natural substances and other known agents.

Engineered: In general, the term “engineered” refers to the aspect ofhaving been manipulated by the hand of man. For example, apolynucleotide is considered to be “engineered” when two or moresequences, that are not linked together in that order in nature, aremanipulated by the hand of man to be directly linked to one another inthe engineered polynucleotide. Comparably, a cell or organism isconsidered to be “engineered” if it has been manipulated so that itsgenetic information is altered (e.g., new genetic material notpreviously present has been introduced, for example by transformation,mating, somatic hybridization, transfection, transduction, or othermechanism, or previously present genetic material is altered or removed,for example by substitution or deletion mutation, or by matingprotocols). As is common practice and is understood by those in the art,progeny of an engineered polynucleotide or cell are typically stillreferred to as “engineered” even though the actual manipulation wasperformed on a prior entity.

Enzyme: As used herein, an “enzyme” is a molecule that catalyzes one ormore biochemical reactions. In some embodiments, an enzyme is orcomprises a polypeptide and/or RNA. In some embodiments, an enzyme is apolypeptide. In some embodiments, an enzyme is a polypeptide and thatranges from about 50 amino acid residues to 2,500 amino acid residues.Enzymes can be classified according to the reaction they catalyze. Insome embodiments, enzymes include oxidoreductases (e.g., catalyzeoxidation/reduction reactions), transferases (e.g., transfer afunctional group, such as, for example, a methyl or phosphate group),hydrolases (e.g., catalyze hydrolysis of various bonds), lyases (e.g.,cleave various bonds by means other than hydrolysis and oxidation),isomerases (e.g., catalyze isomerization changes within a singlemolecule) and ligases (e.g., join two molecules with covalent bonds). Insome embodiments, a member of an enzyme class or family showssignificant sequence homology or identity with, shares a common sequencemotif (e.g., a characteristic sequence element) with, and/or shares acommon activity (in some embodiments at a comparable level or within adesignated range) with a reference enzyme of the class; in someembodiments with all enzymes within the class). For example, in someembodiments, a member enzyme shows an overall degree of sequencehomology or identity with a reference enzyme that is at least about30-40%, and is often greater than about 50%, 60%, 70%, 80%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more and/or includes at leastone region (e.g., a conserved region that may in some embodiments be orcomprise a characteristic sequence element) that shows very highsequence identity, often greater than 90% or even 95%, 96%, 97%, 98%, or99%. Such a conserved region usually encompasses at least 3-4 and oftenup to 20 or more amino acids; in some embodiments, a conserved regionencompasses at least one stretch of at least 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15 or more contiguous amino acids. In some embodiments,an enzyme suitable for use in the context of the present disclosure isof animal origin. In some embodiments, an enzyme suitable for use in thecontext of the present disclosure is of plant origin. In someembodiments, an enzyme suitable for use in the context of the presentdisclosure is of fungal origin. In some embodiments, an enzyme suitablefor use in the context of the present disclosure is of bacterial origin.

Functional: As used herein, a “functional” biological molecule is abiological molecule in a form in which it exhibits a property and/oractivity by which it is characterized.

Homology: As used herein, the term “homology” refers to the overallrelatedness between polymeric molecules, e.g., between nucleic acidmolecules (e.g., DNA molecules and/or RNA molecules) and/or betweenpolypeptide molecules. In some embodiments, polymeric molecules areconsidered to be “homologous” to one another if their sequences are atleast 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95%, or 99% identical. In some embodiments, polymeric molecules areconsidered to be “homologous” to one another if their sequences are atleast 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95%, or 99% similar (e.g., containing residues with relatedchemical properties at corresponding positions).

Human: In some embodiments, a human is an embryo, a fetus, an infant, achild, a teenager, an adult, or a senior citizen.

Identity: As used herein, the term “identity” refers to the overallrelatedness between polymeric molecules, e.g., between nucleic acidmolecules (e.g., DNA molecules and/or RNA molecules) and/or betweenpolypeptide molecules. In some embodiments, polymeric molecules areconsidered to be “substantially identical” to one another if theirsequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,75%, 80%, 85%, 90%, 95%, or 99% identical. Calculation of the percentidentity of two nucleic acid or polypeptide sequences, for example, canbe performed by aligning the two sequences for optimal comparisonpurposes (e.g., gaps can be introduced in one or both of a first and asecond sequences for optimal alignment and non-identical sequences canbe disregarded for comparison purposes). In certain embodiments, thelength of a sequence aligned for comparison purposes is at least 30%, atleast 40%, at least 50%, at least 60%, at least 70%, at least 80%, atleast 90%, at least 95%, or substantially 100% of the length of areference sequence. The nucleotides at corresponding positions are thencompared. When a position in the first sequence is occupied by the sameresidue (e.g., nucleotide or amino acid) as the corresponding positionin the second sequence, then the molecules are identical at thatposition. The percent identity between the two sequences is a functionof the number of identical positions shared by the sequences, takinginto account the number of gaps, and the length of each gap, which needsto be introduced for optimal alignment of the two sequences. Thecomparison of sequences and determination of percent identity betweentwo sequences can be accomplished using a mathematical algorithm.

Polypeptide: As used herein refers to any polymeric chain of aminoacids. In some embodiments, a polypeptide has an amino acid sequencethat occurs in nature. In some embodiments, a polypeptide has an aminoacid sequence that does not occur in nature. In some embodiments, apolypeptide has an amino acid sequence that is engineered in that it isdesigned and/or produced through action of the hand of man. In someembodiments, a polypeptide may comprise or consist of natural aminoacids, non-natural amino acids, or both. In some embodiments, apolypeptide may comprise D-amino acids, L-amino acids, or both. In someembodiments, a polypeptide may include one or more pendant groups orother modifications, e.g., modifying or attached to one or more aminoacid side chains, at the polypeptide's N-terminus, at the polypeptide'sC-terminus, or any combination thereof. In some embodiments, suchpendant groups or modifications may be selected from the groupconsisting of acetylation, amidation, lipidation, methylation,pegylation, etc., including combinations thereof. In some embodiments, auseful polypeptide may comprise or consist of a fragment of a parentpolypeptide. The term “peptide” is generally used to refer to apolypeptide having a length of less than about 100 amino acids, lessthan about 50 amino acids, less than 20 amino acids, or less than 10amino acids.

Recombinant: As used herein, is intended to refer to polypeptides thatare designed, engineered, prepared, expressed, created, manufactured,and/or or isolated by recombinant means, such as polypeptides expressedusing a recombinant expression vector transfected into a host cell;polypeptides isolated from a recombinant, combinatorial humanpolypeptide library; polypeptides isolated from an animal (e.g., amouse, rabbit, sheep, fish, etc.) that is transgenic for or otherwisehas been manipulated to express a gene or genes, or gene components thatencode and/or direct expression of the polypeptide or one or morecomponent(s), portion(s), element(s), or domain(s) thereof; and/orpolypeptides prepared, expressed, created or isolated by any other meansthat involves splicing or ligating selected nucleic acid sequenceelements to one another, chemically synthesizing selected sequenceelements, and/or otherwise generating a nucleic acid that encodes and/ordirects expression of the polypeptide or one or more component(s),portion(s), element(s), or domain(s) thereof In some embodiments, one ormore of such selected sequence elements is found in nature. In someembodiments, one or more of such selected sequence elements is designedin silico. In some embodiments, one or more such selected sequenceelements results from mutagenesis (e.g., in vivo or in vitro) of a knownsequence element, e.g., from a natural or synthetic source.

DETAILED DESCRIPTION

The present disclosure provides methods and compositions that protectcomponents of a composition (e.g., polymers) from damage by lipids(e.g., fatty acids) and/or other compounds that may cause damage. Alsoprovided herein are compositions, methods, systems, and kits thatinclude a polypeptide and/or an enzyme that catalyzes, degrades,modifies and/or sequesters a lipid.

Enzymes

In some embodiments, provided herein are enzymes useful for breakdown ofcompounds (e.g., fatty acids) that may cause damage to components of acomposition (e.g., polymers). In some embodiments, enzymes for use inthe context of the present disclosure include dioxygenases,monooxygenases, heme peroxidases, P450s, and combinations and/orvariants thereof In some embodiments, each of the enzymes can degrade anunsaturated fatty acid. In some embodiments, an enzyme specificallydegrades one or more unsaturated fatty acids.

In some embodiments, compositions and methods of the present disclosurecomprise two or more enzymes that promote breakdown of compounds thatcause damage. In some embodiments, two or more enzymes each promotebreakdown of unsaturated fatty acids.

In some embodiments, compositions and methods of the present disclosurecomprise a plurality of enzymes. In some embodiments, each of theenzymes can degrade an unsaturated fatty acid. In some embodiments, eachof the plurality of enzymes exhibit different substrate specificity forone or more unsaturated fatty acids.

In some embodiments, an enzyme is of animal origin. In some embodiments,an enzyme is of plant origin. In some embodiments, an enzyme is offungal origin. In some embodiments, an enzyme is of bacterial origin. Insome embodiments, an enzyme is a cyanobacterial enzyme. In someembodiments, an enzyme is from archaea.

In some embodiments, an enzyme catalyzes beta or omega oxidation. Insome embodiments, an enzyme catalyzes beta or omega oxidation of fattyacids. In some embodiments, an enzyme has activity at the 5, 8, 9, 10,11, 12, 13 or 15-carbon of a fatty acid. In some embodiments, an enzymehas activity at a 5R, 5S, 8R, 8S, 9R, 9S, 10S, 11R, 11S, 12R, 12S, 13R,13S and/or 15S position of a fatty acid. In some embodiments, an enzymehas activity at the 10S and/or 12S-carbon of a fatty acid (e.g., anoleic acid). In some embodiments, an enzyme for use in the context ofthe present disclosure does not require adenosine triphosphate (ATP) forcatalytic activity.

In some embodiments, a polymeric composition comprises an enzyme withina range from about 0.0001% to about 20% on w/w basis. In someembodiments, a polymeric composition comprises an enzyme within a rangebounded by a lower limit and an upper limit, the upper limit beinglarger than the lower limit. In some embodiments, the lower limit may beabout 0.0001% , 0.0002%, 0.0005%, 0.001%, 0.002%, 0.005%, 0.01%, 0.02%,0.05%, 0.1%, 0.2%, 0.5%, 1%, 2%, 5%, or 10%. In some embodiments, theupper limit may be about 0.0005%, 0.001%, 0.002%, 0.005%, 0.01%, 0.02%,0.05%, 0.1%, 0.2%, 0.5%, 1%, 2%, 5%, 10%, or 20%.

Lipoxygenases

In some embodiments, an enzyme for use in the context of the presentdisclosure is a lipoxygenase. Lipoxygenase (LOX) enzymes catalyzeoxygen-dependent oxidation of fatty acid substrates (for example,linoleic acid and arachidonic acid) to form hydroperoxy-fatty acidproducts. LOX enzymes are categorized as dioxygenases. Certain LOXenzymes have been purified from diverse organisms that display a broadrange of substrate specificity and product specificity (e.g., site ofoxidation within a fatty acid). LOX enzymes are widely expressed inanimals, plants, and fungi, and cyanobacteria.

The present disclosure encompasses the recognition that lipoxygenasesare a suitable class of enzymes for the degradation of unsaturated fattyacids (UFA) such as, for example, oleic acid. Without wishing to bebound by theory, LOX enzymes may convert unsaturated fatty acids tohydroperoxides, which can spontaneously degrade at the site of thedouble bond if not stabilized. In some embodiments, lipoxygenases mayutilize either iron or manganese in their active sites.

In some embodiments, a LOX does not require ATP or other cofactors forcatalytic activity. The present disclosure encompasses the recognitionthat activity independent of ATP or other cofactors is a desirablecharacteristic for certain applications.

In some embodiments, a lipoxygenase suitable for use in the context ofthe present disclosure is of animal origin. In some embodiments, alipoxygenase suitable for use in the context of the present disclosureis of plant origin. In some embodiments, a lipoxygenase suitable for usein the context of the present disclosure is of fungal origin. In someembodiments, a lipoxygenase suitable for use in the context of thepresent disclosure is of bacterial origin. In some embodiments, alipoxygenase suitable for use in the context of the present disclosureis of cyanobacterial origin. In some embodiments, a lipoxygenasesuitable for use in the context of the present disclosure is fromarchaea.

In some embodiments, a LOX can catalyze hydroperoxidation of a fattyacid substrate. In some embodiments, a fatty acid substrate is anunsaturated fatty acid. In some embodiments, a LOX facilitates catalysisof palmitoleic acid, oleic acid, myristoleic acid, linoleic acid, and/orarachidonic acid.

In some embodiments, a LOX facilitates partial or complete degradationof one or more unsaturated fatty acids. In some embodiments, a LOXfacilitates partial or complete degradation of palmitoleic acid, oleicacid, myristoleic acid, linoleic acid, and/or arachidonic acid.

In some embodiments, a LOX has activity (i.e., can catalyzehydroperoxidation) at the 5, 8, 9, 10, 11, 12, 13 or 15-carbon of afatty acid. In some embodiments, a LOX has activity (i.e., can catalyzehydroperoxidation) at a 5R, 5S, 8R, 8S, 9R, 9S, 10S, 11R, 11S, 12R, 12S,13R, 13S and/or 15S position of a fatty acid.

Bioprospecting identified LOX enzymes which act on oleic acid with a 10-or 12-carbon LOX preference. In some embodiments, a lipoxygenasecatalyzes hydroperoxidation at the 10-carbon position of a fatty acid.In some embodiments, a lipoxygenase catalyzes hydroperoxidation at the12- carbon position of a fatty acid. In some embodiments, a LOXcatalyzes hydroperoxidation at the 10S and/or 12S-carbon of a fatty acid(i.e., has 10S-LOX or 12S-LOX activity).

In some embodiments, a lipoxygenase has 10S-LOX activity. In someembodiments, a lipoxygenase is a 10S-LOX from a plant, fungus, bacteria,or archaea. In some embodiments, a 10S-LOX facilitates catalysis ofoleic acid. In some embodiments, a 10S-LOX is from cyanobacteria. Insome embodiments, a 10S-LOX is from cyanobacteria and facilitatescatalysis of oleic acid.

A common source of LOX is from plants (for example soybean), withspecificity for an unsaturated fatty acid in which LOX activity occursat the 9- or 13-carbon. In some embodiments, a lipoxygenase is a9/13-LOX, for example a 9/13-LOX from plants, bacteria, archaea, orfungi.

The following sequences are representative of LOX enzymes that werecharacterized to have 10S-LOX activity.

In some embodiments, a 10S-LOX is or comprises a sequence any one of SEQID NOs: 1 to 12. In some embodiments, a 10S-LOX enzyme has a sequencethat is at least about 50% (e.g., at least about 55%, 60%, 65%, 70%,75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%) identical to anyone of SEQ ID NOs: 1 to 12.

(SEQ ID NO: 1) TRDTSRDGFSNKALAYTLTHFKPIWNLVQSYEPLKRKLNKFFLNSIIYKLPTRPLPYSLMGLDPKIPGTDIPKKTDTYISWDSLTDKTYTGRHLPPDPEFNKEGNLPPLDKVKTLFQKRDGKTIYSEKSTLLFPYWVQWFTDSFLRIDQENRFKNTSNHQIDMCNVYGLTRKQTNMLRAFKDGKFKTQKLKRKDGVEEEYPLFYYADPEQGIIDPQFEGLHAPLNDEKRQPPEKKSKLFAMGVERANVQIGYVMLNTLCIREHNRICDVLSKSYPEWDDERLFQTARNILMVIVLNIIMEEYIFHITPYNFRFFADPEAFTKESWYRENWMAIEFSFVYRWHSAIPETFIYDGKEQSMYDSLWNNQMLIDKGLGALMEETCSQPGTRIGLFNTPDFKIAGTPYTFIDATELASVKLGRQAQLASYNDYREMCGYPRVTDFNQITGDEYAQQKLKELYGHVDKIELFVGLYAEDVRKNSAIPPLVARIIGIDAFSQALTNPLLSPKVFNKETFSEVGWEIIQNTKTVSDLVNRNVPPSDPKYKVSFEL (SEQ ID NO: 2)RDTSKDGFRNKLETYALTHFKPIWNLIQSNDTLKKKVNKFLVNNAIYKVPTRPYPFSTMSPYTSWDSLSDRTYSGLHLPPLDWQPLTNENHLKLKLADTKDFEKKLPAIEDLRGLYRKSGETKYSPKSTLIFPYFVQWFTDSFLRTDRHNHRKNTSNHQIDLCTVYGLNAKITHLLRSYQGGKLKSQIINGEEYPPFYYDEKGEAKKEFIGLPHQLDNDGNPKADTFPLDKKQKLFAMGVEVERSNVQIGYVMLNVLALREHNRLCELLAKTYPSWDDERLFQTARNILIVEVLRIVVEDYVNHITPYHFQFITDPLTFSNEKWYRQNWMTVEFTLVYRWHSMLPDTLIYNGQKIPTYETQWNNEMIIKQGLGALFEESCSQPCAQLSLFNTPEFLIPVELASVRFGREVKLRSYNDYRQLCKYPRVTDFDQISSDKNIQKELQRLYGHVDNIELYVGIYAEDLRENSALPSLVGRLIGIDAFSQVLTNPLLAESVFHPETFSPVGWEEIQNTKTLSQLLHRNLPPSDKKYRVSFDRAST (SEQ ID NO: 3)AGKRDTSKDGFDNKVQTFLLTNFKGIWEIVQSNEFLKRKVNKTLINSLIYKIPTRPNPYSMMTLDEYIPDTKIPKKTDTYTSWELLNDRTYIGRHLPPDPKFNSEGNLPKVEDLAVLFRKRDGKTIYSPKSTMLFPYWVQWFTDSFLRIDHTKEKKLKNTSNHEIDLCNVYGLNRKRTHLLRTFKGGKFKTQKLKRQDGIEEEYPLFYYADPAQGIVDPQFDGLYEPINDEKRLPADKKQYLFAMGVERANVQIGYVMLNTLCIREHNRLCDELASNYPDWDDERLFQTSRNILMAIILNIIMEEYINHITPYHFKLFADPAAFVKESWYRPNWMTIEFDFVYRWHSAIPETFIYDGQPTDIAASLWNNKMFIDKGLGALMEETCSQPGTRIGLFNTPDILVELTELPSIRLGRQLQLASYNDYREMCGFPRVTKFEQITGDEFAQEKLKELYGHVDNIEFYVGLYAEEVRKNSTIPPLVARLIGIDAFSEALNNPLLSPTIFNKDTFSPVGWEIIQNTKTVSDLINRNVPPSDKKYKVTFDL (SEQ ID NO: 4)TRDTSRDGFSNKALAYTLTHFKPIWNLVQSYEPLKRKLNKFFLNSIIYKLPTRPLPYSLMGLDPKIPGTDIPKKTDTYISWDSLTDKTYTGRHLPPDPEFNKEGNLPPLDKVKTLFQKRDGKTIYSEKSTLLFPYWVQWFTDSFLRIDQENRFKNTSNHQIDMCNVYGLTRKQTNMLRAFKDGKFKTQKLKRKDGVEEEYPLFYYADPEQGIIDPQFEGLHAPLNDEKRQPPEKKSKLFAMGVERANVQIGYVMLNTLCIREHNRICDVLSKSYPEWDDERLFQTARNILMVIVLNIIMEEYIFHITPYNFRFFADPEAFTKESWYRENWMAIEFSFVYRWHSAIPETFIYDGKEQSMYDSLWNNQMLIDKGLGALMEETCSQPGTRIGLFNTPDFKIAGTPYTFIDATELASVKLGRQAQLASYNDYREMCGYPRVTDFNQITGDEYAQQKLKELYGHVDKIELFVGLYAEDVRKNSAIPPLVARIIGIDAFSQALTNPLLSPKVFNKETFSEVGWEIIQNTKTVSDLVNRNVPPSDPKYKVSFEE (SEQ ID NO: 5)RDTSKDGFRNKLETYALTHFKPIWNLIQSNDTLKKKVNKFLVNNAIYKVPTRPYPFSTMSPYTSWDSLSDRTYSGLHLPPLDWQPLTNENHLKLKLADTKDFEKKLPAIEDLRGLYRKSGETKYSPKSTLIFPYFVQWFTDSFLRTDRHNHRKNTSNHQIDLCTVYGLNAKITHLLRSYQGGKLKSQIINGEEYPPFYYDEKGEAKKEFIGLPHQLDNDGNPKADTFPLDKKQKLFAMGVEVERSNVQIGYVMLNVLALREHNRLCELLAKTYPSWDDERLFQTARNILIVEVLRIVVEDYVNHITPYHFQFITDPLTFSNEKWYRQNWMTVEFTLVYRWHSMLPDTLIYNGQKIPTYETQWNNEMIIKQGLGALFEESCSQPCAQLSLFNTPEFLIPVELASVRFGREVKLRSYNDYRQLCKYPRVTDFDQISSDKNIQKELQRLYGHVDNIELYVGIYAEDLRENSALPSLVGRLIGIDAFSQVLTNPLLAESVFHPETFSPVGWEEIQNTKTLSQLLHRNLPPSDKKYRVSFDRASE (SEQ ID NO: 6)AGKRDTSKDGFDNKVQTFLLTNFKGIWEIVQSNEFLKRKVNKTLINSLIYKIPTRPNPYSMMTLDEYIPDTKIPKKTDTYTSWELLNDRTYIGRHLPPDPKFNSEGNLPKVEDLAVLFRKRDGKTIYSPKSTMLFPYWVQWFTDSFLRIDHTKEKKLKNTSNHEIDLCNVYGLNRKRTHLLRTFKGGKFKTQKLKRQDGIEEEYPLFYYADPAQGIVDPQFDGLYEPINDEKRLPADKKQYLFAMGVERANVQIGYVMLNTLCIREHNRLCDELASNYPDWDDERLFQTSRNILMAIILNIIMEEYINHITPYHFKLFADPAAFVKESWYRPNWMTIEFDFVYRWHSAIPETFIYDGQPTDIAASLWNNKMFIDKGLGALMEETCSQPGTRIGLFNTPDILVELTELPSIRLGRQLQLASYNDYREMCGFPRVTKFEQITGDEFAQEKLKELYGHVDNIEFYVGLYAEEVRKNSTIPPLVARLIGIDAFSEALNNPLLSPTIFNKDTFSPVGWEIIQNTKTVSDLINRNVPPSDKKYKVTFDE (SEQ ID NO: 7)TRDTSRDGFSNKALAYTLTHFKPIWNLVQSYEPLKRKLNKFFLNSIIYKLPTRPLPYSLMGLDPKIPGTDIPKKTDTYISWDSLTDKTYTGRHLPPDPEFNKEGNLPPLDKVKTLFQKRDGKTIYSEKSTLLFPYWVQWFTDSFLRIDQENRFKNTSNHQIDMCNVYGLTRKQTNMLRAFKDGKFKTQKLKRKDGVEEEYPLFYYADPEQGIIDPQFEGLHAPLNDEKRQPPEKKSKLFAMGVERANVQIGYVMLNTLCIREHNRICDVLSKSYPEWDDERLFQTARNILMVIVLNIIMEEYIFHITPYNFRFFADPEAFTKESWYRENWMAIEFSFVYRWHSAIPETFIYDGKEQSMYDSLWNNQMLIDKGLGALMEETCSQPGTRIGLFNTPDFKIAGTPYTFIDATELASVKLGRQAQLASYNDYREMCGYPRVTDFNQITGDEYAQQKLKELYGHVDKIELFVGLYAEDVRKNSAIPPLVARIIGIDAFSQALTNPLLSPKVFNKETFSEVGWEIIQNTKTVSDLVNRNVPPSDPKYKVSFED (SEQ ID NO: 8)RDTSKDGFRNKLETYALTHFKPIWNLIQSNDTLKKKVNKFLVNNAIYKVPTRPYPFSTMSPYTSWDSLSDRTYSGLHLPPLDWQPLTNENHLKLKLADTKDFEKKLPAIEDLRGLYRKSGETKYSPKSTLIFPYFVQWFTDSFLRTDRHNHRKNTSNHQIDLCTVYGLNAKITHLLRSYQGGKLKSQIINGEEYPPFYYDEKGEAKKEFIGLPHQLDNDGNPKADTFPLDKKQKLFAMGVEVERSNVQIGYVMLNVLALREHNRLCELLAKTYPSWDDERLFQTARNILIVEVLRIVVEDYVNHITPYHFQFITDPLTFSNEKWYRQNWMTVEFTLVYRWHSMLPDTLIYNGQKIPTYETQWNNEMIIKQGLGALFEESCSQPCAQLSLFNTPEFLIPVELASVRFGREVKLRSYNDYRQLCKYPRVTDFDQISSDKNIQKELQRLYGHVDNIELYVGIYAEDLRENSALPSLVGRLIGIDAFSQVLTNPLLAESVFHPETFSPVGWEEIQNTKTLSQLLHRNLPPSDKKYRVSFDRASD (SEQ ID NO: 9)AGKRDTSKDGFDNKVQTFLLTNFKGIWEIVQSNEFLKRKVNKTLINSLIYKIPTRPNPYSMMTLDEYIPDTKIPKKTDTYTSWELLNDRTYIGRHLPPDPKFNSEGNLPKVEDLAVLFRKRDGKTIYSPKSTMLFPYWVQWFTDSFLRIDHTKEKKLKNTSNHEIDLCNVYGLNRKRTHLLRTFKGGKFKTQKLKRQDGIEEEYPLFYYADPAQGIVDPQFDGLYEPINDEKRLPADKKQYLFAMGVERANVQIGYVMLNTLCIREHNRLCDELASNYPDWDDERLFQTSRNILMAIILNIIMEEYINHITPYHFKLFADPAAFVKESWYRPNWMTIEFDFVYRWHSAIPETFIYDGQPTDIAASLWNNKMFIDKGLGALMEETCSQPGTRIGLFNTPDILVELTELPSIRLGRQLQLASYNDYREMCGFPRVTKFEQITGDEFAQEKLKELYGHVDNIEFYVGLYAEEVRKNSTIPPLVARLIGIDAFSEALNNPLLSPTIFNKDTFSPVGWEIIQNTKTVSDLINRNVPPSDKKYKVTFDD (SEQ ID NO: 10)TRDTSRDGFSNKALAYTLTHFKPIWNLVQSYEPLKRKLNKFFLNSIIYKLPTRPLPYSLMGLDPKIPGTDIPKKTDTYISWDSLTDKTYTGRHLPPDPEFNKEGNLPPLDKVKTLFQKRDGKTIYSEKSTLLFPYWVQWFTDSFLRIDQENRFKNTSNHQIDMCNVYGLTRKQTNMLRAFKDGKFKTQKLKRKDGVEEEYPLFYYADPEQGIIDPQFEGLHAPLNDEKRQPPEKKSKLFAMGVERANVQIGYVMLNTLCIREHNRICDVLSKSYPEWDDERLFQTARNILMVIVLNIIMEEYIFHITPYNFRFFADPEAFTKESWYRENWMAIEFSFVYRWHSAIPETFIYDGKEQSMYDSLWNNQMLIDKGLGALMEETCSQPGTRIGLFNTPDFKIAGTPYTFIDATELASVKLGRQAQLASYNDYREMCGYPRVTDFNQITGDEYAQQKLKELYGHVDKIELFVGLYAEDVRKNSAIPPLVARIIGIDAFSQALTNPLLSPKVFNKETFSEVGWEIIQNTKTVSDLVNRNVPPSDPKYKVSFET (SEQ ID NO: 11)RDTSKDGFRNKLETYALTHFKPIWNLIQSNDTLKKKVNKFLVNNAIYKVPTRPYPFSTMSPYTSWDSLSDRTYSGLHLPPLDWQPLTNENHLKLKLADTKDFEKKLPAIEDLRGLYRKSGETKYSPKSTLIFPYFVQWFTDSFLRTDRHNHRKNTSNHQIDLCTVYGLNAKITHLLRSYQGGKLKSQIINGEEYPPFYYDEKGEAKKEFIGLPHQLDNDGNPKADTFPLDKKQKLFAMGVEVERSNVQIGYVMLNVLALREHNRLCELLAKTYPSWDDERLFQTARNILIVEVLRIVVEDYVNHITPYHFQFITDPLTFSNEKWYRQNWMTVEFTLVYRWHSMLPDTLIYNGQKIPTYETQWNNEMIIKQGLGALFEESCSQPCAQLSLFNTPEFLIPVELASVRFGREVKLRSYNDYRQLCKYPRVTDFDQISSDKNIQKELQRLYGHVDNIELYVGIYAEDLRENSALPSLVGRLIGIDAFSQVLTNPLLAESVFHPETFSPVGWEEIQNTKTLSQLLHRNLPPSDKKYRVSFDRASL (SEQ ID NO: 12)AGKRDTSKDGFDNKVQTFLLTNFKGIWEIVQSNEFLKRKVNKTLINSLIYKIPTRPNPYSMMTLDEYIPDTKIPKKTDTYTSWELLNDRTYIGRHLPPDPKFNSEGNLPKVEDLAVLFRKRDGKTIYSPKSTMLFPYWVQWFTDSFLRIDHTKEKKLKNTSNHEIDLCNVYGLNRKRTHLLRTFKGGKFKTQKLKRQDGIEEEYPLFYYADPAQGIVDPQFDGLYEPINDEKRLPADKKQYLFAMGVERANVQIGYVMLNTLCIREHNRLCDELASNYPDWDDERLFQTSRNILMAIILNIIMEEYINHITPYHFKLFADPAAFVKESWYRPNWMTIEFDFVYRWHSAIPETFIYDGQPTDIAASLWNNKMFIDKGLGALMEETCSQPGTRIGLFNTPDILVELTELPSIRLGRQLQLASYNDYREMCGFPRVTKFEQITGDEFAQEKLKELYGHVDNIEFYVGLYAEEVRKNSTIPPLVARLIGIDAFSEALNNPLLSPTIFNKDTFSPVGWEIIQNTKTVSDLINRNVPPSDK KYKVTFDT

Also provided are herein are circular permutated variants of any enzymeor polypeptide described herein. In circular permutation methods, the N-and C-terminus of the mutated protein are redefined by geneticengineering. Circular permutation methods have been described in theart. For example, Graf and Schachmann, (Proc. Natl. Acad. Sci. USA(1996) 93, 11591-11596), provide an overview of enzymes and otherproteins with which circular permutation methods have been carried out.Methods for engineering circular permutated polypeptide (e.g., enzyme)variants are known in the art, see, e.g., US Pat. Publ. No.2010/0196991. In some embodiments, a circular permutation variantincludes a linker.

In some embodiments, an enzyme for use in the context of the presentdisclosure is a cyanobacterial enzyme with LOX activity (e.g., 10S-LOXactivity) or variant thereof. In some embodiments, a cyanobacterialprostaglandin-endoperoxide synthase enzyme or variant thereof has LOXactivity (e.g., 10S-LOX activity). In some embodiments, an enzyme withLOX catalytic activity (e.g., 10S-LOX activity) comprises an amino acidsequence of SEQ ID NO: 13 and/or a circular permutation thereof.

(SEQ ID NO: 13) TRDTSRDGFSNKALAYTLTHFKPIWNLVQSYEPLKRKLNKFFLNSIIYKLPTRPLPYSLMGLDPKIPGTDIPKKTDTYISWDSLTDKTYTGRHLPPDPEFNKEGNLPPLDKVKTLFQKRDGKTIYSEKSTLLFPYWVQWFTDSFLRIDQENRFKNTSNHQIDMCNVYGLTRKQTNMLRAFKDGKFKTQKLKRKDGVEEEYPLFYYADPEQGIIDPQFEGLHAPLNDEKRQPPEKKSKLFAMGVERANVQIGYVMLNTLCIREHNRICDVLSKSYPEWDDERLFQTARNILMVIVLNIIMEEYIFHITPYNFRFFADPEAFTKESWYRENWMAIEFSFVYRWHSAIPETFIYDGKEQSMYDSLWNNQMLIDKGLGALMEETCSQPGTRIGLFNTPDFKIAGTPYTFIDATELASVKLGRQAQLASYNDYREMCGYPRVTDFNQITGDEYAQQKLKELYGHVDKIELFVGLYAEDVRKNSAIPPLVARIIGIDAFSQALTNPLLSPKVFNKETFSEVGWEIIQNTKTVSDLVNRNVPPSDPKYKVSFEX,where X is any amino acid.

In some embodiments, a cyanobacterial heme peroxide synthase enzyme orvariant thereof has LOX activity (e.g., 10S-LOX activity). In someembodiments, an enzyme with LOX (e.g., 10S-LOX activity) catalyticactivity comprises an amino acid sequence of SEQ ID NO: 14 and/or acircular permutation thereof. In some embodiments, an enzyme with LOX(e.g., 10S-LOX activity) catalytic activity comprises an amino acidsequence of SEQ ID NO: 15 and/or a circular permutation thereof.

(SEQ ID NO: 14) RDTSKDGFRNKLETYALTHFKPIWNLIQSNDTLKKKVNKFLVNNAIYKVPTRPYPFSTMSPYTSWDSLSDRTYSGLHLPPLDWQPLTNENHLKLKLADTKDFEKKLPAIEDLRGLYRKSGETKYSPKSTLIFPYFVQWFTDSFLRTDRHNHRKNTSNHQIDLCTVYGLNAKITHLLRSYQGGKLKSQIINGEEYPPFYYDEKGEAKKEFIGLPHQLDNDGNPKADTFPLDKKQKLFAMGVEVERSNVQIGYVMLNVLALREHNRLCELLAKTYPSWDDERLFQTARNILIVEVLRIVVEDYVNHITPYHFQFITDPLTFSNEKWYRQNWMTVEFTLVYRWHSMLPDTLIYNGQKIPTYETQWNNEMIIKQGLGALFEESCSQPCAQLSLFNTPEFLIPVELASVRFGREVKLRSYNDYRQLCKYPRVTDFDQISSDKNIQKELQRLYGHVDNIELYVGIYAEDLRENSALPSLVGRLIGIDAFSQVLTNPLLAESVFHPETFSPVGWEEIQNTKTLSQLLHRNLPPSDKKYRVSFDRASX, where X is any amino acid.(SEQ ID NO: 15) AGKRDTSKDGFDNKVQTFLLTNFKGIWEIVQSNEFLKRKVNKTLINSLIYKIPTRPNPYSMMTLDEYIPDTKIPKKTDTYTSWELLNDRTYIGRHLPPDPKFNSEGNLPKVEDLAVLFRKRDGKTIYSPKSTMLFPYWVQWFTDSFLRIDHTKEKKLKNTSNHEIDLCNVYGLNRKRTHLLRTFKGGKFKTQKLKRQDGIEEEYPLFYYADPAQGIVDPQFDGLYEPINDEKRLPADKKQYLFAMGVERANVQIGYVMLNTLCIREHNRLCDELASNYPDWDDERLFQTSRNILMAIILNIIMEEYINHITPYHFKLFADPAAFVKESWYRPNWMTIEFDFVYRWHSAIPETFIYDGQPTDIAASLWNNKMFIDKGLGALMEETCSQPGTRIGLFNTPDILVELTELPSIRLGRQLQLASYNDYREMCGFPRVTKFEQITGDEFAQEKLKELYGHVDNIEFYVGLYAEEVRKNSTIPPLVARLIGIDAFSEALNNPLLSPTIFNKDTFSPVGWEIIQNTKTVSDLINRNVPPSDKKYKVTFDLX, where X is any amino acid.

A sequence similarity network of LOX enzymes is tested for activity onoleic acid and depicted in FIG. 5. Gray nodes indicate 10S-LOX activity.These sequences are distinct from 9/13-LOX from plants, bacteria, andfungi, and are sometimes annotated as heme peroxidases. In particular,the cluster contains primarily cyanobacterial enzymes contains the10S-LOXs that are active on oleic acid.

In some embodiments, a fatty acid to be degraded is oleic acid, and aLOX enzyme specific for action on the 10S- or 12S-carbon of oleic acidis employed in a composition or method of the present disclosure.

Dioxygenases and Monooxygenases

In some embodiments, dioxygenases other than LOX, as well asmonooxygenases, are suitable for use in the context of the presentdisclosure. In some embodiments, an enzyme is a dioxygenase. In someembodiments, an enzyme is a monooxygenase.

In some embodiments, a dioxygenase catalyzes oxidation of a fatty acidsubstrate. In some embodiments, a fatty acid substrate is an unsaturatedfatty acid. In some embodiments, a dioxygenase catalyzes oxidation ofpalmitoleic acid, oleic acid, myristoleic acid, linoleic acid, and/orarachidonic acid.

In some embodiments, a dioxygenase catalyzes the partial or completedegradation of one or more unsaturated fatty acids. In some embodiments,a dioxygenase facilitates partial or complete degradation of palmitoleicacid, oleic acid, myristoleic acid, linoleic acid, and/or arachidonicacid.

In some embodiments, a monooxygenase catalyzes oxidation of a fatty acidsubstrate. In some embodiments, a fatty acid substrate is an unsaturatedfatty acid. In some embodiments, a monooxygenase catalyzes oxidation ofpalmitoleic acid, oleic acid, myristoleic acid, linoleic acid, and/orarachidonic acid.

In some embodiments, a monooxygenase catalyzes the partial or completedegradation of one or more unsaturated fatty acids. In some embodiments,a monooxygenase facilitates partial or complete degradation palmitoleicacid, oleic acid, myristoleic acid, linoleic acid, and/or arachidonicacid.

In some embodiments, a dioxygenase and/or a monooxygenase is from aplant, bacteria, archaea, or fungus. In some embodiments, a dioxygenaseand/or a monooxygenase facilitates catalysis of oleic acid. In someembodiments, a dioxygenase and/or a monooxygenase is from cyanobacteria.In some embodiments, a dioxygenase and/or a monooxygenase is fromcyanobacteria and facilitates catalysis of oleic acid.

In some embodiments, an enzyme catalyzes beta or omega oxidation. Insome embodiments, an enzyme catalyzes oxidation or hydroperoxidation atthe 5, 8, 9, 10, 11, 12, 13 or 15-carbon of a fatty acid. In someembodiments, an enzyme catalyzes oxidation or hydroperoxidation ata 5R,5S, 8R, 8S, 9R, 9S, 10S, 11R, 11S, 12R, 12S, 13R, 13S and/or 15Sposition of a fatty acid. In some embodiments, an enzyme catalyzesoxidation or hydroperoxidation of the 10S and/or 12S-carbon of a fattyacid (e.g., an oleic acid). In some embodiments, an enzyme for use inthe context of the present disclosure does not require adenosinetriphosphate (ATP) for catalytic activity.

In some embodiments, a monooxygenase and/or dioxygenase in the contextof the present disclosure can catalyze partial or complete degradationof a fatty acid (e.g., an unsaturated fatty acid). In some embodiments,activity of a monooxygenase and/or dioxygenase may be independent ofcofactors (e.g., ATP). In some embodiments, activity of a monooxygenaseand/or dioxygenase may require a cofactor (e.g., ATP).

In some embodiments, compositions and methods of the present disclosurecomprise a LOX and one or more other dioxygenases that promote breakdownof compounds that cause damage to polymers. In some embodiments,compositions and methods of the present disclosure comprise a LOX andone or more monooxygenases that promote breakdown of compounds thatcause damage to polymers. In some embodiments, compositions and methodsof the present disclosure comprise a plurality of enzymes. In someembodiments, each of the enzymes can degrade an unsaturated fatty acid.In some embodiments, each of the plurality of enzymes exhibit differentsubstrate specificity for one or more unsaturated fatty acids.

P450s

In some embodiments, an enzyme suitable for use in the context of thepresent disclosure is a P450. Cytochrome P450 enzymes form a superfamilyof hemoproteins found in bacteria, archaea and eukaryotes. In a commonactivity, cytochrome P450 acts as a monooxygenase, by inserting oneoxygen atom of molecular oxygen into a substrate molecule, while theother oxygen atom is reduced to water. A P450 catalytic reaction mayrequire two electrons for the activation of molecular oxygen. P450s fromeukaryotes use NADPH as the external reductant and source of electrons.Each electron may be transferred one at a time to a cytochrome P450active site. In some embodiments, an electron transfer may be donated byan electron donor protein, e.g., a cytochrome P450 reductase (CPR). ACPR may be an electron donor protein for several different P450s fromthe same or from different organisms. In some cases P450s can also becoupled to a cytochrome b5 protein that can act as the electron donorprotein or can improve the efficiency of the electron transfer from theCPR to the P450. In eukaryotic cells and particularly in plants, P450sand CPRs are generally membrane-bound proteins and are associated withthe endoplasmic reticulum. These proteins may be anchored to themembrane by an N-terminal trans-membrane helix.

Many P450s have low substrate specificity and are therefore able tocatalyze the oxidation of many diverse structures. Many P450s have aparticular region and stereo-selectivity with a given substrate; howeverthey produce a mixture of several products from a particular substrate.In some embodiments, a P450 is involved in breakdown and detoxificationof molecules (e.g., xenobiotics). In some embodiments, a P450 isinvolved in a biosynthetic pathway. P450s involved in a biosyntheticpathway may exhibit specificity for certain types of substrates andregion and stereo-selectivity. In some embodiments, a P450 is from aplant, bacteria, or fungus.

A large number of P450s can be found in nature and particularly inplants. One plant genome can contain several hundreds of genes encodingfor P450s.

In some embodiments, a P450 is active on one or more unsaturated fattyacids. In some embodiments, a P450 facilitates catalysis of palmitoleicacid, oleic acid, myristoleic acid, linoleic acid, and/or arachidonicacid. In some embodiments, a P450 facilitates catalysis of oleic acid.

In some embodiments, compositions and methods of the present disclosurecomprise a P450 and another enzyme (e.g., a dioxygenase, monooxygenase,heme peroxidase) that promote breakdown of compounds that cause damage.In some embodiments, compositions and methods of the present disclosurecomprise a plurality of enzymes. In some embodiments, each of theenzymes can act on an unsaturated fatty acid. In some embodiments, eachof the enzymes can degrade an unsaturated fatty acid. In someembodiments, each of the plurality of enzymes exhibit differentsubstrate specificity for one or more unsaturated fatty acids.

Heme Peroxidases

In some embodiments, an enzyme for use in the context of the presentdisclosure is a heme peroxidase. In some embodiments, a heme peroxidasehas a ferriprotoporphyrin IX prosthetic group located at the activesite. The plant enzymes horseradish peroxidase (HRP) and plant soyabeanperoxidase (SBP) are examples of plant heme peroxidases.

In some embodiments, a heme peroxidase facilitates catalysis of one ormore unsaturated fatty acids. In some embodiments, a heme peroxidasefacilitates catalysis of palmitoleic acid, oleic acid, myristoleic acid,linoleic acid, and/or arachidonic acid. In some embodiments, a hemeperoxidase facilitates catalysis of oleic acid.

In some embodiments, compositions and methods of the present disclosurecomprise a heme peroxidase and another enzyme (e.g., a dioxygenase,monooxygenase, P450) that promotes breakdown of compounds that causedamage to polymers. In some embodiments, compositions and methods of thepresent disclosure comprise a plurality of enzymes. In some embodiments,each of the enzymes facilitates catalysis of an unsaturated fatty acid.In some embodiments, each of the enzymes can degrade an unsaturatedfatty acid. In some embodiments, each of the plurality of enzymesexhibit different substrate specificity for one or more unsaturatedfatty acids.

Polypeptides

In some embodiments, protection of materials from damaging lipids (e.g.,fatty acids, such as oleic acid and other unsaturated fatty acids), isadditionally or alternatively performed via non-catalytic binding of aprotein to a lipid. In some embodiments, binding to a lipid (e.g., anunsaturated fatty acid) results in formation of a protein-lipid complexthat would be less able to diffuse into the material being protected.Binding of a lipid (e.g., an unsaturated fatty acid) could also limit orprevent it from reacting with other materials, such as a polymericmaterial or other material to be protected.

In some embodiments, compositions are provided comprising one or moreenzymes that promote breakdown of a fatty acid and one or morepolypeptides that form a complex with a fatty acid.

Nucleic Acid Construction and Expression

Enzymes and polypeptides as described herein may be produced fromnucleic acid molecules using molecular biological methods known to theart. Nucleic acid molecules are inserted into a vector that is able toexpress the polypeptide when introduced into an appropriate host cell.Appropriate host cells include, but are not limited to, bacterial,yeast, insect, and mammalian cells. Any of the methods known to oneskilled in the art for the insertion of DNA fragments into a vector maybe used to construct expression vectors encoding the enzymes of thepresent disclosure under control of transcriptional/translationalcontrol signals. These methods may include in vitro recombinant DNA andsynthetic techniques and in vivo recombination (See Sambrook et al.,Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory;Current Protocols in Molecular Biology, Eds. Ausubel, et al, GreenePubl. Assoc., Wiley-Interscience, N.Y.).

Expression of nucleic acid molecules in accordance with the presentdisclosure may be regulated by a second nucleic acid sequence so thatthe molecule is expressed in a host transformed with the recombinant DNAmolecule. For example, expression of the nucleic acid molecules of thepresent disclosure may be controlled by promoter and/or enhancerelements, which are known in the art.

Nucleic acid constructs of the present disclosure may be inserted intoan expression vector or viral vector by methods known to the art, andnucleic acid molecules operatively linked to an expression controlsequence.

Where appropriate, nucleic acid sequences that encode enzyme asdescribed herein may be modified to include codons that are optimizedfor expression in a particular cell type or organism. Codon optimizedsequences are synthetic sequences, and preferably encode the identicalpolypeptide (or a biologically active fragment of a full lengthpolypeptide which has substantially the same activity as the full lengthpolypeptide) encoded by the non-codon optimized parent polynucleotide.In some embodiments, the coding region of the genetic material encodingantibody components, in whole or in part, may include an alteredsequence to optimize codon usage for a particular cell type (e.g., aeukaryotic or prokaryotic cell). For example, the coding sequence for anenzyme as described herein may be optimized for expression in bacterialcells, fungal cells, plant cells, mammalian cells, etc. Such a sequencemay be described as a codon-optimized sequence.

An expression vector containing a nucleic acid molecule is transformedinto a suitable host cell to allow for production of the protein encodedby the nucleic acid constructs. Exemplary host cells include prokaryotes(e.g., E. coli) and eukaryotes (e.g., yeast cells or mammalian cells).Host cells transformed with an expression vector are grown underconditions permitting production of an enzyme of the present disclosurefollowed by recovery of the enzyme or polypeptide.

Enzyme and polypeptides of the present disclosure may be purified by anytechnique, which allows for the subsequent formation of a stable enzyme.For example, not wishing to be bound by theory, enzymes may be recoveredfrom cells either as soluble polypeptides or as inclusion bodies, fromwhich they may be extracted quantitatively by 8 M guanidiniumhydrochloride and dialysis. In order to further purify enzymes of thepresent disclosure, conventional ion exchange chromatography,hydrophobic interaction chromatography, reverse phase chromatography orgel filtration may be used. Enzymes of the present invention may also berecovered from conditioned media following secretion from eukaryotic orprokaryotic cells.

Polymers

Biological fluids may include components that can damage polymers. Forexample, fatty acids (e.g., unsaturated fatty acids or other fattyacids) may include reactive compounds such as peroxide that can damagepolymers. The present invention encompasses the recognition thatprotective enzymes may be incorporated into a polymer to prevent,mitigate or lessen damage to a polymer and/or a device containing thepolymer. The protective enzymes may include any enzymes described hereinsuitable to protect against biological substances (e.g., fatty acids)that may degrade or otherwise damage polymeric materials.

In some embodiments, polymers suitable for use in the context of thepresent disclosure may include polyamides, polyesters,polyaryletherketones, polyimides, polyetherimides, polyamideimide,liquid crystalline polymers, polycarbonates, polyolefins, polyphenyleneoxide, polysulfones, polyacrylates, acrylonitrile butadiene styrenepolymer, polyoxymethylene, polystyrene, polyarylene sulfide,polyvinylidene fluoride, polytetrafluoroethylene, polyvinylidenechloride, polyvinyl chloride, and any other suitable polymer. In someembodiments, a polymer is or comprises a silicone resin, epoxy resin,polyvinyl butyral resin, cellulose acetate, ethylene-vinyl acetatecopolymer (EVA) or an ionomer. In some embodiments, a polymer is orcomprises an acrylic-based polymer. In some embodiments, a polymer is orcomprises a silicon-based polymer.

In some embodiments, a protective enzyme or polypeptide is embeddedwithin a polymeric composition. In some embodiments, a polymericcomposition comprises a protective enzyme or polypeptide within a rangefrom about 0.0001% to about 20% on w/w basis. In some embodiments, apolymeric composition comprises two or more protective enzymes, eachpresent within a range from about 0.0001% to about 20% on w/w basis. Insome embodiments, a polymeric composition comprises a protective enzymeor polypeptide within a range bounded by a lower limit and an upperlimit, the upper limit being larger than the lower limit. In someembodiments, the lower limit may be about 0.0001% , 0.0002%, 0.0005%,0.001%, 0.002%, 0.005%, 0.01%, 0.02%, 0.05%, 0.1%, 0.2%, 0.5%, 1%, 2%,5%, or 10%. In some embodiments, the upper limit may be about 0.0005%,0.001%, 0.002%, 0.005%, 0.01%, 0.02%, 0.05%, 0.1%, 0.2%, 0.5%, 1%, 2%,5%, 10%, or 20%.

In some embodiments, a protective enzyme is applied to the surface of apolymeric composition. In some embodiments, a polymeric composition iscoated with a protective enzyme.

Components of Electronic Devices

Electronic devices may be exposed to the bodily fluids of users such assweat and sebum. These bodily fluids may contain fatty acids thatdegrade polymers in electronic devices. To prevent degradation,protective enzymes may be incorporated into one or more components ofelectronic device. Protective enzymes may include, for example, one ormore enzymes from the lipoxygenase (LOX) enzyme family, or othersuitable enzymes that degrade harmful substances such as fatty acids.Protective enzymes such as lipoxygenase enzymes can degrade thesereactive compounds (e.g., peroxide) and thereby reduce the ability of afatty acid (e.g., oleic acid, etc.) to degrade polymers and damageelectronic devices. Protective enzymes may neutralize the destructiveactivity of the fatty acids and thereby help enhance robustness ofpolymer structures for electronic devices.

In some embodiments, a polymer with a protective enzyme may be part ofan adhesive, gasket, tape, button, or other structure that may bevulnerable to damage by exposure to biological lipids (e.g., fattyacids). A protective enzyme may be incorporated into a coating, anadhesive, a gasket, or other structures in an electronic device. Whenfatty acids come into contact with a protective enzyme in a coating,adhesive, gasket, or other structure, the fatty acids in the structuresare neutralized.

In general, any suitable components in an electronic device may includeone or more protective enzymes (e.g., dioxygenases, monooxygenases, hemeperoxidases, P450s, and/or lipoxygenases). For example, a protectiveenzyme may be incorporated into plastic portions of housings, gaskets,adhesive layers, tape layers, coatings, gap filling sealant and othersealants, liquid polymers that are dispensed as coatings, roomtemperature adhesives, fluoropolymer coatings and/or other hydrophobiccoatings, liquid polymer materials that serve as carrier fluids forenzyme dispensing without serving as structural adhesive, and/or othermaterials (e.g., polymers) in an electronic device.

Topical Formulations

In some embodiments, a protective enzyme and/or polypeptide may be usedas part of a topical formulation for application to the skin of a mammal(e.g., a human). In some embodiments, a protective enzyme and/orpolypeptide may be used as part of a topical formulation for applicationto a polymer. In some embodiments, the polymer is a component of adevice. In some embodiments, a protective enzyme and/or polypeptide maybe used as part of a topical formulation for application to a devicethat comprises a polymer. Without wishing to be bound by theory, it isenvisioned that inclusion of a protective enzyme and/or polypeptide mayprevent degradation of other components of a topical formulation. Insome embodiments, two or more protective enzymes may be used a part of atopical formulation for application to the skin of a mammal (e.g., ahuman). In some embodiments a mixture of enzymes may be used as part ofa topical formulation. For example, it may be desirable to combineenzymes with specificities to multiple unsaturated fatty acids, forexample to both oleic and linoleic acid present in sebum and sweat.Similarly, single enzymes or mixtures could be employed to target oleicacid or multiple unsaturated fatty acids in products applied to theskin.

In some embodiments, a topical formulation comprising a protectiveenzyme and/or polypeptide is an emulsion, gel, ointment, or lotion.Topical formulations may be prepared using methods known in the art, forexample, as provided by reference texts such as, REMINGTON: THE SCIENCEAND PRACTICE OF PHARMACY 1577-1591, 1672-1673, 866-885; (Alfonso R.Gennaro ed. 19th ed. 1995); Ghosh, T. K.; et al. TRANSDERMAL AND TOPICALDRUG DELIVERY SYSTEMS (1997), both of which are hereby incorporatedherein by reference.

In some embodiments, a protective enzyme and/or polypeptide may beuseful for compositions comprising a medicament for topical formulation.In some embodiments, a protective enzyme and/or polypeptide is acomponent of a topical sunscreen formulation.

In some embodiments, a topical formulation comprises a protective enzymeor polypeptide within a range from about 0.0001% to about 20% on w/wbasis. In some embodiments, a polymeric composition comprises two ormore protective enzymes, each present within a range from about 0.0001%to about 20% on w/w basis. In some embodiments, a topical formulationcomprises a protective enzyme or polypeptide within a range bounded by alower limit and an upper limit, the upper limit being larger than thelower limit. In some embodiments, the lower limit may be about 0.0001% ,0.0002%, 0.0005%, 0.001%, 0.002%, 0.005%, 0.01%, 0.02%, 0.05%, 0.1%,0.2%, 0.5%, 1%, 2%, 5%, or 10%. In some embodiments, the upper limit maybe about 0.0005%, 0.001%, 0.002%, 0.005%, 0.01%, 0.02%, 0.05%, 0.1%,0.2%, 0.5%, 1%, 2%, 5%, 10%, or 20%.

The foregoing is merely illustrative and various modifications can bemade to the described embodiments. The foregoing embodiments may beimplemented individually or in any combination.

EXAMPLES Example 1—An Exemplary Protective Enzyme Prevents Fatty AcidInduced Polymer Swelling

This example shows the enzymatic activity of a series of LOX enzymesthat were assayed for activity on oleic acid. The chemical activity ofthe enzymes that is measured is shown in FIG. 1. The assay detects the10-HOME product derived from treatment of the unstable peroxide producedby the LOX enzyme with the reducing agent DTT. Enzymes were tested inthe context of E. coli cell lysate. E. coli overexpressing the LOXenzyme were induced, then cells were lysed enzymatically using lysozyme.The lysate was added to the reaction containing oleic acid, dodecylmaltoside as a surfactant and Tris buffer pH 7.6 and the reaction wasrun at room temperature for 15 minutes. The reaction product was reducedusing DTT and quenched in a methanol/acetonitrile mixture.and 10-HOMEproduct was quantified using LC-MS. Resulting activity is shown in FIG.2.

Example 2—An Exemplary Protective Enzyme Prevents Fatty Acid InducedPolymer Swelling

This example demonstrates the efficacy of an exemplary protective enzyme(e.g., a lipoxygenase). Specifically, this Example demonstrates that apolymer composition comprising an exemplary protective enzyme (e.g., alipoxygenase) is able to prevent polymer swelling induced by treatmentwith a fatty acid. An acrylic foam pressure sensitive adhesive tape (3M™VHB™ Tape 4914-015) was used as a representative composition to assessthe efficacy protective enzymes to prevent polymeric swelling. Thisrepresentative adhesive was exposed to an exemplary unsaturated fattyacid (oleic acid), and polymer swelling was assessed over time for bothlipoxygenase treated (treated) and untreated samples, FIG. 3. Y-axisindicates the amount of swelling by area, X-axis indicates the number ofhours post-exposure. As can be seen in FIG. 3, beginning atapproximately 40 hours, the untreated samples show an increase inswelling greater than that of the treated samples. Thus, treatment witha lipoxygenase protected the representative adhesive from fatty acidinduced damage over time.

Example 3—An Exemplary of an alternative assay demonstrating ProtectiveEnzyme Efficacy

This example provides additional evidence of the protective effect of aprotective enzyme on the swelling induced in an adhesive by fatty acids.An acrylic foam pressure sensitive adhesive tape (3M™ VHB™ Tape4914-015) was used as a representative composition to assess theefficacy protective enzymes to prevent polymeric swelling. Thisrepresentative adhesive was first treated with either lipoxygenase orleft untreated and then exposed to an exemplary unsaturated fatty acid(oleic acid). Polymer swelling was assessed over time for bothlipoxygenase treated (treated) and untreated samples, FIG. 4. Y-axisindicates the amount of swelling by area, X-axis indicates the number ofhours post-exposure. As can be seen in FIG. 4, the untreated samplesshow an increase in swelling greater than that of the treated samples.Thus, pre-treatment with a lipoxygenase protected the representativeadhesive from fatty acid induced damage over time.

Example 4—Analysis of LOX family Enzymes for Activity

This example describes assessment of a network of LOX enzymes forcatalytic activity on an exemplary fatty acid substrate (e.g., oleicacid). Homologs of extant lipoxygenases were identified viabioinformatics methods, based on sequence and structural similarity.Each node represents a candidate LOX enzyme, with each enzyme clusteredby pairwise similarity to the other enzymes in the library. Gray nodeswere shown to have 10S-LOX activity on oleic acid. These 10S-LOX enzymesare distantly related to the more well-described 9S/13S-LOX enzymesdescribed for plants and bacteria. The above example demonstrates thatlipoxygenases are a class of enzyme that can degrade an exemplary fattyacid, oleic acid. Moreover, it is determined that other enzymes, such asdioxygenases, monooxygenases, heme peroxidases, P450s, and others couldalso catalyze the degradation of lipids such as fatty acids.

Enzyme family networks are depicted in FIG. 5, with gray nodesindicating possession 10S-LOX activity. Thus, several different hemeperoxidase enzymes possess 10S-LOX activity. These sequences aredistinct from 9/13-LOX from plants, bacteria, and fungi. Of note, thecluster contains primarily cyanobacterial enzymes was found to contain10S-LOXs that are active on the specific exemplary fatty acid (oleicacid). The assays described herein can be used to characterize theability of enzymes to protect from damage induced by lipids (e.g.,specific fatty acids and/or combinations of fatty acids).

EXEMPLARY EMBODIMENTS

1. A method for promoting the breakdown of one or more fatty acids inbodily fluid secreted from a human, the method comprising contacting thebodily fluid with an enzyme.

2. A method for inhibiting swelling of a polymer caused by exposure to afatty acid, the method comprising contacting the polymer with an enzyme.

3. A method for inhibiting swelling of a polymer caused by exposure to afatty acid, the method comprising contacting the polymer with apolypeptide that binds a fatty acid.

4. The method according to any one of embodiments 1-3 wherein the fattyacid is unsaturated.

5. The method according to any one of embodiments 1-3 wherein the fattyacid is an oleic acid.

6. The method according to any one of embodiments 1-3 wherein the fattyacid is a linoleic acid.

7. The method according to any one of embodiments 1-6 wherein the enzymeor polypeptide is embedded in a polymer.

8. The method according to any one of embodiments 1 or 4-7 wherein thebodily fluid comprises sweat.

9. The method according to any one of embodiments 1 or 4-7 wherein thebodily fluid comprises sebum.

10. The method according to any one of embodiments 1-2, or 4-9 whereinthe enzyme is a dioxygenase.

11. The method according to any one of embodiments 1-2, or 4-9 whereinthe enzyme is a lipoxygenase.

12. The method according to any one of embodiments 1-2, or 4-9 whereinthe enzyme is a monooxygenase.

13. The method according to any one of embodiments 1-2, or 4-9 whereinthe enzyme does not require adenosine triphosphate (ATP) for catalyticactivity.

14. The method according to any one of embodiments 1-2, or 4-13comprising contacting the bodily fluid or polymer with a plurality ofenzymes.

15. The method according to embodiment 14, wherein the plurality ofenzymes comprises a dioxygenase, monooxygenase, heme peroxidases, and/orP450s.

16. The method according to any one of embodiments 1-2, or 4-15 whereinthe enzyme is a cyanobacterial enzyme.

17. The method according to any one of embodiments 1-2, or 14-16 whereinthe enzyme catalyzes beta or omega oxidation.

18. The method according to any one of embodiments 1-2, or 14-17 whereinthe enzyme has activity at the 10S and/or 12S-carbon of an oleic acid.

19. The method according to embodiment 3 wherein the polypeptide forms acomplex with the fatty acid.

20. The method according to embodiment 3 wherein the polypeptideinhibits reactivity of the fatty acid.

21. The method according to embodiment 3 wherein the polypeptideinhibits diffusion of the fatty acid.

22. A composition comprising a polymer and one or more enzymes thatpromote breakdown of a fatty acid.

23. A composition comprising a polymer and one or more polypeptides thatbind to a fatty acid.

24. The composition according to embodiment 22 or 23 wherein the one ormore enzymes or polypeptides are embedded in the polymer.

25. The composition according to any one of embodiments 22-24 whereinthe fatty acid is unsaturated.

26. The composition according to any one of embodiments 22-24 whereinthe fatty acid is an oleic acid.

27. The composition according to any one of embodiments 24-24 whereinthe fatty acid is a linoleic acid.

28. The composition according to any one of embodiments 22, or 24-27wherein the enzyme is a dioxygenase.

29. The composition according to any one of embodiments 22, or 24-27wherein the enzyme is a lipoxygenase.

30. The composition according to any one of embodiments 22, or 24-27wherein the enzyme is a monooxygenase.

31. The composition according to any one of embodiments 22, or 24-30wherein the enzyme does not require adenosine triphosphate (ATP) forcatalytic activity.

32. The composition according to any one of embodiments 22, or 24-31comprising a plurality of enzymes.

33. The composition according to embodiment 32 wherein the plurality ofenzymes comprises a dioxygenase, monooxygenase, heme peroxidase, and/orP450.

34. The composition according to any one of embodiments 22, or 25-33wherein the enzyme is a cyanobacterial enzyme.

35. The composition according to any one of embodiments 22, or 25-33wherein the enzyme catalyzes beta or omega oxidation.

36. The composition according to any one of embodiments 22, or 25-33wherein the enzyme has activity at the 10S and/or 12S-carbon of an oleicacid.

37. The composition according to embodiment 23 wherein the polypeptideforms a complex with the fatty acid.

38. The composition according to embodiment 23 wherein the polypeptideinhibits reactivity of the fatty acid.

39. The composition according to embodiment 23 wherein the polypeptideinhibits diffusion of the fatty acid.

40. A composition comprising one or more enzymes according to any one ofembodiments 22, or 25-33 and one or more polypeptides according to anyone of embodiments 23, or 37-39.

41. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of SEQ ID NO:1 or a circular permutated variant thereof..

42. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of SEQ ID NO:2 or a circular permutated variant thereof.

43. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of SEQ ID NO:3 or a circular permutated variant thereof.

44. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of SEQ ID NO:4 or a circular permutated variant thereof.

45. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of SEQ ID NO:5 or a circular permutated variant thereof.

46. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of SEQ ID NO:6 or a circular permutated variant thereof.

47. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of SEQ ID NO:7 or a circular permutated variant thereof.

48. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of SEQ ID NO:8 or a circular permutated variant thereof.

48. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of SEQ ID NO:9 or a circular permutated variant thereof.

49. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of SEQ ID NO:10 or a circular permutated variant thereof.

50. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of SEQ ID NO:11 or a circular permutated variant thereof.

51. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of SEQ ID NO:12 or a circular permutated variant thereof.

52. The method or composition according to any one of embodiments 1-2,or 22 wherein the enzyme comprises an amino acid sequence of any one ofSEQ ID NOs: 13-15, or a circular permutated variant thereof.

Having thus described at least several aspects and embodiments of thisinvention, it is to be appreciated that various alterations,modifications, and improvements will readily be apparent to thoseskilled in the art. Such alterations, modifications, and improvementsare intended to be part of this disclosure, and are intended to bewithin the spirit and scope of the invention. Accordingly, the foregoingdescription and drawings are by way of example only and the invention isdescribed in further detail by the claims that follow.

EQUIVALENTS

The articles “a” and “an” as used herein in the specification and in theclaims, unless clearly indicated to the contrary, should be understoodto include the plural referents. Claims or descriptions that include“or” between one or more members of a group are considered satisfied ifone, more than one, or all of the group members are present in, employedin, or otherwise relevant to a given product or process unless indicatedto the contrary or otherwise evident from the context. The inventionincludes embodiments in which exactly one member of the group is presentin, employed in, or otherwise relevant to a given product or process.The invention also includes embodiments in which more than one, or theentire group members are present in, employed in, or otherwise relevantto a given product or process. Furthermore, it is to be understood thatthe invention encompasses all variations, combinations, and permutationsin which one or more limitations, elements, clauses, descriptive terms,etc., from one or more of the listed claims is introduced into anotherclaim dependent on the same base claim (or, as relevant, any otherclaim) unless otherwise indicated or unless it would be evident to oneof ordinary skill in the art that a contradiction or inconsistency wouldarise. Where elements are presented as lists, (e.g., in Markush group orsimilar format) it is to be understood that each subgroup of theelements is also disclosed, and any element(s) can be removed from thegroup. It should be understood that, in general, where the invention, oraspects of the invention, is/are referred to as comprising particularelements, features, etc., certain embodiments of the invention oraspects of the invention consist, or consist essentially of, suchelements, features, etc. For purposes of simplicity those embodimentshave not in every case been specifically set forth in so many wordsherein. It should also be understood that any embodiment or aspect ofthe invention can be explicitly excluded from the claims, regardless ofwhether the specific exclusion is recited in the specification.

1-34. (canceled)
 35. A host cell that comprises a heterologouspolynucleotide encoding a lipoxygenase, wherein the lipoxygenasecomprises an amino acid sequence that is at least 90% identical to SEQID NO:
 2. 36. The host cell of claim 35, wherein the lipoxygenasecomprises SEQ ID NO:
 2. 37. The host cell of claim 35, wherein thelipoxygenase has activity at the 10S-carbon of one or more fatty acids.38. The host cell of claim 35, wherein the lipoxygenase promotesbreakdown of one or more fatty acids.
 39. The host cell of claim 38,wherein the lipoxygenase is capable of promoting the breakdown of one ormore fatty acids in bodily fluid secreted from a human.
 40. The hostcell of claim 39, wherein the bodily fluid comprises sweat or sebum. 41.The host cell of claim 38, wherein the one or more fatty acids is anoleic acid and/or a linoleic acid.
 42. The host cell of claim 35,wherein the host cell is a plant cell, a fungal cell, a yeast cell, abacterial cell, or an animal cell.
 43. The host cell of claim 42,wherein the bacterial cell is an Escherichia coli cell.
 44. A methodcomprising culturing the host cell of claim
 35. 45. A host cell thatcomprises a heterologous polynucleotide encoding a lipoxygenase, whereinthe lipoxygenase comprises an amino acid sequence that is at least 90%identical to SEQ ID NO:
 3. 46. The host cell of claim 45, wherein thelipoxygenase comprises SEQ ID NO:
 3. 47. The host cell of claim 45,wherein the lipoxygenase has activity at the 10S-carbon of one or morefatty acids.
 48. The host cell of claim 45, wherein the lipoxygenasepromotes breakdown of one or more fatty acids.
 49. The host cell ofclaim 48, wherein the lipoxygenase is capable of promoting the breakdownof one or more fatty acids in bodily fluid secreted from a human. 50.The host cell of claim 49, wherein the bodily fluid comprises sweat orsebum.
 51. The host cell of claim 48, wherein the one or more fattyacids is an oleic acid and/or a linoleic acid.
 52. The host cell ofclaim 45, wherein the host cell is a plant cell, a fungal cell, a yeastcell, a bacterial cell, or an animal cell.
 53. The host cell of claim52, wherein the bacterial cell is an Escherichia coli cell.
 54. A methodcomprising culturing the host cell of claim 45.